Richards C S, Watkins S C, Hoffman E P, Schneider N R, Milsark I W, Katz K S, Cook J D, Kunkel L M, Cortada J M
GeneScreen, Department of Pediatrics, University of Texas Southwestern Medical Center, Dallas 75207.
Am J Hum Genet. 1990 Apr;46(4):672-81.
One of female MZ twins presented with muscular dystrophy. Physical examination, creatine phosphokinase levels, and muscle biopsy were consistent with Duchenne muscular dystrophy (DMD). However, because of her sex she was diagnosed as having limb-girdle muscular dystrophy. With cDNA probes to the DMD gene, a gene deletion was detected in the twins and their mother. The de novo mutation which arose in the mother was shown by novel junction fragments generated by HindIII, PstI, or TaqI when probed with cDNA8. Additional evidence of a large gene deletion was given by novel SfiI junction fragments detected by probes p20, J-Bir, and J-66 on pulsed-field gel electrophoresis (PFGE). Immunoblot analysis of muscle from the affected twin showed dystrophin of normal size but of reduced amount. Immunofluorescent visualization of dystrophin revealed foci of dystrophin-positive fibers adjacent to foci of dystrophin-negative fibers. These data indicate that the affected twin is a manifesting carrier of an abnormal DMD gene, her myopathy being a direct result of underexpression of dystrophin. Cytogenetic analysis revealed normal karyotypes, eliminating the possibility of a translocation affecting DMD gene function. Both linkage analysis and DNA fingerprint analysis revealed that each twin has two different X chromosomes, eliminating the possibility of uniparental disomy as a mechanism for DMD expression. On the basis of methylation differences of the paternal and maternal X chromosomes in these MZ twins, we propose uneven lyonization (X chromosome inactivation) as the underlying mechanism for disease expression in the affected female.
一对女性单卵双胞胎中的一人患有肌肉萎缩症。体格检查、肌酸磷酸激酶水平及肌肉活检结果均与杜兴氏肌肉萎缩症(DMD)相符。然而,由于她的性别,她被诊断为肢带型肌肉萎缩症。使用针对DMD基因的cDNA探针,在这对双胞胎及其母亲中检测到了一个基因缺失。母亲发生的新发突变通过用cDNA8探针检测时由HindIII、PstI或TaqI产生的新连接片段得以显示。通过脉冲场凝胶电泳(PFGE)用探针p20、J - Bir和J - 66检测到的新SfiI连接片段,提供了大基因缺失的额外证据。对患病双胞胎的肌肉进行免疫印迹分析显示,抗肌萎缩蛋白大小正常但数量减少。抗肌萎缩蛋白的免疫荧光可视化显示,抗肌萎缩蛋白阳性纤维灶与抗肌萎缩蛋白阴性纤维灶相邻。这些数据表明,患病的双胞胎是异常DMD基因的显性携带者,她的肌病是抗肌萎缩蛋白表达不足的直接结果。细胞遗传学分析显示核型正常,排除了影响DMD基因功能的易位可能性。连锁分析和DNA指纹分析均显示,每个双胞胎都有两条不同的X染色体,排除了单亲二倍体作为DMD表达机制的可能性。基于这些单卵双胞胎中父本和母本X染色体的甲基化差异,我们提出不均衡X染色体失活(X染色体随机失活)是患病女性疾病表达的潜在机制。